1a-phthaloylglotamylmitomycin c



May 13, 1969 4 Filed Nov. 25, 1964 MASANAO MATSUI ET AL La-PHTHALOYLGLUTAIIYLMITOMYCIN 0 Sheet 046 F/GT/ ATTORNEY wvsmon;

May 1969 MASANAO MATSUI ET AL 3,444,182

LO.PHTHALOYLGLUTAMYLMITOMYCIN C Filed Nov. 25, 1964 A KBQQfiEEMQ:

ooh 8m 009 8: 82 8: 08m 08m 8% 80 9 ON on 04 on 8 oh om 8 OE WQ on: 83 8: 88 8mm 8mm ooow ATTORNEY y 13, 1969 I MASANAO MATSUI E L 4,182

Ln.PHTHALOYLGLUTAMYLMITOMYCIN C Filed Nov. 25, 1964 Sheet a Q I i 4/ INVENTQR 4 ewe ATTORNEY May 13, 1969 "N IASANAO MATSUI E 3,444,132

"Ls-PHTHALOYLGLUTAMYLMITOMYEFEN C Filed Nov- 25. 1964 Sheet i of a IOOOO IOOOO- BY M 44 4016,;

ATTORNEY! May 13, 1969 MASANAO MATSUI E AL 3,444,182

La-PHTHALOYLGLUTAMYLMITOMYCIN 0 Filed Nov. 25, 1964 Sheet 6 01 s 3 g m i Q5 k g LI g s ww @QQ'QS'QW BYMMM X aMZM ATTORNEYS United States Patent 3,444,182 la-PHTHALOYLGLOTAMYLMITOMYCIN C Masanao Matsui, Tokyo, Keizo Uzn, Shizuoka-ken, and

Yasuhiro Yamada and Shigetoshi Wakaki, Tokyo, Japan, assignors to Kyowa Hakko Kogyo Co., Ltd., Tokyo, Japan, a corporation of Japan Filed Nov. 25, 1964, Ser. No. 413,743 Int. Cl. C07d 57/00, 27/00; A6lk 21/00 US. Cl. 260326 1 Claim ABSTRACT OF THE DISCLOSURE The present invention relates to new antibiotics, more particularly, relates to new antibiotics derived from mitomycin C.

Mitomycin C is a well-known antibiotic having the formula:

CHzO C ONH:

OCH;

The present inventors have conducted research in this field and have found several types of derivatives. In Canadian application No. 904,353, now Canadian Patent No. 743,105, granted Sept. 20, 1966, the present inventors have provided a type of derivatives of mitomycin C which has the general formula:

wherein R is alkyl radical.

In the present specification, the term alkyl means not only alkyl but also substituted alkyl.

Several specific derivatives are exemplified in the application. The above derivatives are synthesized by reacting the reagent having alkanoyl-group such as acylchloride and carbonic acid anhydride with mitomycin C.

The present inventors have conducted further studies on the synthesis of derivatives of mitomycin C, and have found 3 specific derivatives belonging to the above-mentioned type of derivatives. Furthermore, the present inventors have found the new types of derivatives; one is obtained by treating mitomycin C with acid, and the other is the la-sulfonyl derivative obtained by reacting the sulfonating agent with mitomycin C.

3,444,182 Patented May 13, 1969 One object of the present invention is to provide new derivatives of mitomycin C, and another object is to provide a process for the synthesis of the derivatives. A further object of the present invention is to provide less poisonous derivatives of mitomycin C. Other objects will easily be understood from the description.

According to the present invention, three specific derivatives belonging to la-acyl derivatives, are 1a-acetyl-, 1achloracetyland la-phthaloyl glutamyl-derivatives of mitomycin C, which are synthesized by reacting the corresponding acylchloride or carbonic acid anhydride with mitomycin C in preferable solvent such as water, methanol, acetone, pyridine, tetrahydrofuran, dimethylformamide and etc. in the presence of deacidifying substances such as inorganic alkali, organic bases (pyridine triethylamine). When no deacidifying substance exists in the reaction mixture, mitomycin C is decomposed by the produced acid and the yield of la-acyl derivative is decreased. However, this phenomenon relates to one of the new derivative of the present invention, i.e. acid-decomposed mitomcyin C. As the chemical property of mitomycin C, it has been known that mitomycin C is easily decomposed by acid, however, the decomposed product has been neither realized nor isolated. The present inventors have studied this decomposition and isolated then identified the product. The decomposition is shown by the following formula:

CHaOCONH:

The derivative (I) is, however, further decomposed as follows:

0 Hm I CHIOCONHQ 11,0

I N 0H 0 HO CHQOCONHQ HO l 1 11,0 rno N on g l N OH NHz NH:

therefore, it is necessary for obtaining the derivative (I 1-hydroxy-2,7-diaminomitoseneat high yield to select the favorable conditions of acidity and reaction time, for example, when the concentration of hydrochloric acid is 0.1 N, the favourable reaction time is 10-15 min., while when the concentration is 0.005 N, the favourable reaction time is about 20 hours. As to the acid used in the decomposing reaction, inorganic acid such as hydrochloric, sulfuric, phosphoric acids and etc. and also organic acid such as acetic, citric acids and etc. may be used. The decomposition reaction is preferably carried out in suitable solvents such as water, methanol, acetone, tetrahydrofuran. The recovery of the product is favourably carried out by using chromatographic method as described in examples of the present specification.

Another type of derivatives of the present invention is la-sulfonyl derivatives of mitomycin C which are synthesized by reacting sulfonating reagent with mitomycin C in the preferable solvent aforementioned in the presence of deacidifying substances in order to prevent the acid decomposition. The la-sulfonyl derivative is shown as follows:

Example 1.1a-acetylmitomycin C HzN CHzO C ONH:

'OCHQ ((CHaC)zO) 1 g. of mitomycin C was dissolved in ml. of pyridine and 2 ml. of acetic anhydride was added on stirring. Standing for 30 min. at room temperature, the reaction mixture was concentrated in vacuo. The resulting pastelike concentrate was dissolved in chloroform and the chromatographic treatment was carried out using alumina. The upper-most band was developed and eluated by 5% methanol-containing chloroform. The eluate was concentrated in vacuo, and the resulting concentrate was dissolved in acetone. Adding petroleum ether, brownish purple amorphous precipitate of la-acetylmitomycin C was obtained. Yield 800 mg.

Analysis..Calcd. for C H O N (CH CO); C, 54.25; H, 5.36; N, 14.89; OH CO, 11. 6. Found: C, 53.70; H, 5.34; N, 13.80; CH CO, 12.14.

The ultraviolet adsorption in ethanol is shown in FIG. 1, and

xgg 215 mu =24,300) and 258 mu (e 19,800)

The infrared spectrum in Nujol is shown in FIG. 2.

Example 2.--1a-monochloroacetylmitomycin C HzN CHzOC ONHr OCH; 0 ClCHzC 0 C1 N A ((ClCH2CO)20) NCOCH2C1 xgfi 214 m (e =24,500) and 358 my (e 18,500)

The infrared spectrum in Nujol is shown in FIG. 4.

Example 3.1a-phthaloylglutamylmitomycin C I omocoNm um OCH:

N-CHG O OC C O 1 g. of mitomycin C and 0.8 g. of phthaloylglutamic acid anhydride were dissolved in 20 m1. of pyridine. Standing for 4 days at room temperature, the reaction mixture was concentrated in vacuo and the resulting concentrate dissolved in MeOH:CHCl (1:3). The chromatographic treatment carried out using silicic acid. 1.5 g. of the brownish amorphous powder of la-phthaloylglutamylmitomycin C was obtained.

Analysis.Calcd. for C28 H27O10N5: C, H, N, 11.80. Found: C, 55.66; H, 5.22; N, 11.01.

The ultraviolet adsorption in ethanol is shown in FIG. 5.

1%,? 218 mp (e saooo) and 357 mp (e 3300) The infrared spectrum in Nujol is shown in FIG. 6.

Example 4.-1a-methanesulfonylmitomycin C CHaSOzCl 1 g. of mitomycin C was dissolved in 20 ml. of pyridine and added with 400 mg. of triethylamine. To this solution, 350 mg. of methanesulfonylchloride in 5 ml. of benzene was added dropwise on stirring and cooling. Stirring for still 20 min., the precipitated triethylaminehydrochloride was filtered off, and the filtrate was concentrated. The resulting paste-like residue was dissolved in ethylacetate. The chromatographic treatment was carried out using silicic acid. The band was developed and eluated by acetonezethylacetate (1:2), whereby the product and unreacted mitomycin C were separated. The first eluate was concentrated and added with petroleum ether. The precipitate was separated and dried. 850 mg. of purple amorphous powder of la-methane sulfonylmitomycin C was obtained.

Analysis.Calcd. for C H O N S: C, 46.59; H, 4.88; N, 11.16. Found: C, 46.35; H, 5.08; N, 11.50.

The ultraviolet adsorption in ethanol is shown in FIG. 7,

215 mp (e =22,000) and 356 mp The infrared spectrum in Nujol is shown in FIG. 8.

umQQ-Smpm 1a-p-toluenesulfonylmitomycin C was prepared by the same process as in Example 4 except using p-toluenesulfonic acid chloride in place of methanesulfonylchloride.

Analysis.-Calcd. for C H O N S: C, 54.08; H, 4.95; N, 11.47. Found: C, 53.85; H, 4.86; N, 11.07.

The ultraviolet adsorption in ethanol is shown in FIG. 9,

(32H OH max.

216 m (e =24,000) and 354 mp The infrared spectrum in Nujol is shown in FIG. 10.

Example 6.-1-hydroxy-2,7-diaminomitosene CHzO C O NH:

H2N CHzO C ONH:

1 g. of mitomycin C was dissolved in 0.1 N hydrochloric acid. After standing for 15 min. at room temperature, the reddish purple solution was adjusted to be pH 9 by adding aqueous ammonia. The solution was kept in the refrigerator and the resultant precipitate was separated and washed with a small amount of acetone. 850 mg. of crude 1-hydroxy-2,7-diaminomitosene was obtained.

The crude product was dissolved in acetone and adsorbed on silicic acid in a column. The development was carried out by using acetone as the solvent and chromatogram of 3 bands was observed. The elution was carried out by using also acetone as solvent and the first eluate was concentrated in vacuo. The resultant dark reddish needle crystal of 1-hydroxy-2,7-diaminomitosene was separated. Yield: 550 mg.

Analysis.-Calcd. for C H O N C, 52.49; H, 5.04; N, 17.49. Found: C, 52.35; H, 5.20; N, 17.30.

The ultraviolet adsorption in ethanol is shown in FIG. 11,

xggf 248 m (e 11,800)

=19,100) and 508 m (e 1 1 What we claim is: 1. The chemical compound 1a-phthalaylglutamylitomycin '0 having the formula:

References Cited UNITED STATES PATENTS 3,226,393 12/1965 Meyer et a1. 2603263 XR 20 3,179,671 4/1965 Mowat et a1. 260319 1 2 OTHER REFERENCES Burger: Medicinal Chemistry, page 43 (1960), RS. 403B8.

Skorokhodov et a1.: Chemical Abstracts, vol. 57, col. 8555 (1962).

Elderfield: Heterocyclic Compounds (1950), QD 400E4, p. 71, vol. 1.

Noller: Chemistry of Organic Compounds (1957), QD 253N65, p. 10 4.

Wagner et a1.: Synthetic Organic Chemistry (1953),

Webb et al.: J. Am. Chem. Soc., vol. 84, pp 3185-88 QD 262W24, pp. 566, 82223. (1962).

Tulinsky, 1.: Am. Chem. Soc., vol. 84, p. 3188 (1962).

ALEX MAZEL, Primary Examiner.

J. A. NARCAVAGE, Assistant Examiner.

US. Cl. X.R. 2603263, 999 

1. THE CHEMICAL COMPOUND LA-PHTHALOYLGLUTAMYLITOMYCIN C HAVING THE FORMULA: 